Cell lysis

Cell lysis (for western, IP, etc.)

1. Lyse cells on ice using 0.5-1ml EB for 5 min. then spin at 20 000g for 5 min. at 4C.

2. Take the supernatant and mix 1/1 with 2x protein sample buffer, heat for 5 min. at 95C.

3. Store at -80C.

    0.5 M NaCl
    10 mM EDTA, pH 8
    1% Triton X100
    20 mM Tris-Cl, pH 7.0
    20mM NaF
    20mM glycerophosphate (optional)
    Just before using add: 2 mM sodium pervanadate (heat and mix before adding)
          1 mM PMSF
          30 ug aprotinin

2x sample buffer:
    125 mM Tris Base
    20% (v/v) Glycerol
    2% (w/v) SDS
    0.02% Bromophenol blue
    2% mecaptoethanol

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